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Genotyping common SNP and a microsatellite sequence closely linked to waxy gene in rice by DNA based markers | ||
Iranian Journal of Genetics and Plant Breeding | ||
مقاله 1، دوره 5، شماره 1 - شماره پیاپی 9، تیر 2016، صفحه 1-7 اصل مقاله (712.58 K) | ||
شناسه دیجیتال (DOI): 10.30479/ijgpb.2016.1038 | ||
نویسندگان | ||
mojtaba najafi* ؛ Ghodrat Rahimi؛ Zarbakht Ansari؛ Mohammad Ali Rouhi | ||
Department of Animal Science, Sari Agriculture Sciences and Natural Resources University, P. O. Box: 48181-68984, Sari, Iran. | ||
تاریخ دریافت: 06 اردیبهشت 1396، تاریخ پذیرش: 06 اردیبهشت 1396 | ||
چکیده | ||
The potential of different DNA based molecular markers was examined for the detection of single nucleotide polymorphism (SNP) in the waxy gene and a microsatellite (SSR) sequence closely linked to it in a collection of rice varieties. DNA was extracted from leaf samples of 68 different rice cultivars by the CTAB method and specific primers were designed for the amplification of waxy gene and SSR marker loci. After amplification of the desired fragments by polymerase chain reaction (PCR) genotyping of samples was carried out by the melting curve analysis technique. To confirm the results of the melting curve genotyping, two methods of PCR-SSCP and PBR were used. The presence of two genotypes (TT and GG) in the waxy gene observed from these two methods was perfectly consistent with the results of the melting curve analysis. The frequency of each TT and GG genotype was estimated 25 and 75 percent, respectively. Nine different genotypes including BH, HH, FF, DD, CC, EE, BB, AA and EG were found in the SSR marker site. This indicates the existence of a high genetic variability in the studied varieties. Although, there were no associations between the G-T polymorphism with the traits, the association of different patterns in SSR marker with the GW, TGW and FGN traits was significant. The results showed that the melting curve analysis technique is a fast and inexpensive method that can be used for polymorphism detection. Morever, this SSR marker site can be used as a suitable marker for detecting some economical traits in rice breeding. | ||
کلیدواژهها | ||
Melting curve analysis؛ Microsatellite marker؛ PBR؛ PCR-SSCP | ||
عنوان مقاله [English] | ||
بررسی چند شکلی تک نوکلئوتیدی و توالی های ریزماهواره ای در واریته های مختلف برنج با استفاده از نشانگرهای مبتنی بر DNA | ||
نویسندگان [English] | ||
مجتبی نجفی؛ قدرت رحیمی؛ زربخت انصاری؛ محمد علی روحی | ||
دانشگاه علوم کشاورزی و منابع طبیعی ساری، دانشکده علوم دامی و شیلات، آزمایشگاه ژنتیک مولکولی دام، ایران، کدپستی: 48986-18184. | ||
چکیده [English] | ||
در پژوهش حاضر، نشانگرهای مولکولی مختلف مبتنی بر DNA برای شناسایی چند شکلی تک نوکلئوتیدی و توالیهای میکروساتالایت برای ژن واکسی در واریتههای برنج مورد بررسی قرار گرفت. استخراج DNA از نمونههای برگی 68 رقم برنج با استفاده از روش CTAB انجام گرفت و پرایمرهای اختصاصی برای تکثیر ژن واکسی طراحی گردید. بعد از تکثیر قطعات مورد نظر با استفاده از PCR، توسط تکنیک منحنی ذوب، تعیین ژنوتیپ صورت گرفت. برای تایید نتایج تعیین ژنوتیپ با آنالیز منحنی ذوب دو روش PBR و PCR-SSCP برای جایگاه ژن واکسی مورد استفاده قرار گرفتند. نتایج دو ژنوتیپ TT و GG را در ژن واکسی نشان داد که با نتایج بدست آمده از دو روش PBR و PCR-SSCP کاملا مطابقت داشت. فراوانی ژنوتیپهای TT و GG به ترتیب 25 و 75 درصد برآورد شد. هم چنین، نه ژنوتیپ مختلف شامل BH، HH، FF، DD، CC، EE، BB، AA و EG در نشانگر SSR شناسایی شد که نشان دهنده تنوع ژنتیکی بالا در واریتههای مختلف برنج مورد مطالعه میباشد. اگرچه همبستگی چند شکلی G-T با صفات معنی دار نشد، ولی همبستگی معنی داری بین ژنوتیپهای مشاهده شده در نشانگر SSR با صفات GW، TGW و FGN به دست آمد. نتایج این مطالعه نشان داد که تکنیک آنالیز منحنی ذوب به عنوان یک روش سریع و کارآمد و ارزان میتواند به منظور تعیین ژنوتیپ مورد استفاده قرار گیرد. هم چنین، نتایج حاصله از نشانگر SSR نشان داد این نشانگر یک نشانگر کارآمدی برای برخی از صفات اقتصادی در به نژادی واریتههای برنج میباشد. | ||
کلیدواژهها [English] | ||
آنالیز منحنی ذوب, نشانگر ریزماهواره ای, PBR, PCR-SSCP | ||
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